EXPERIMENTAL'

Sp-Hydroxy-19-nor-5a- androstane- i'i+-o?&?- p - toluenesulfo- nate (I). A 17-mg. sample of 3p-hydroxy-19-nor-5a-andro- stan-17-one (m.p. 177-179°)6 was dissolved in 2.0 ml. dry pyridine containing 500 mg. of freshly recrystallized p- toluenesulfonyl chloride.7 The solution was allowed to stand a t room temperature for 24 hr. About 15 ml. of ice water was added and the resulting suspension extracted with cold chloroform. The chloroform phase was washed with cold 0.2,V hydrochloric acid, cold 5% aqueous sodium bicarbon- ate and cold water till neutral, dried over sodium sulfate, and evaporated under reduced pressure to dryness. A 28.2- mg. sample of Rolid resulted (I); XE: 5.78 (cyclopentyl C=O), 6.25 (phenyl C=C) 7.4, 8.5, and 14.95 p; no hy- droxyl absorption wan present. A similar spectrum was ob- tained with the tosylate of epiandrosterone. Sa-Hydroxy-19-nor-6a-androstan-l7-one (11) f r o m (I).

The crude tosylate ( I ) was dissolved in 4.0 ml. of dimethyl- formamide containing 180 mg. of potassium acetate in 0.5 ml. of wat.er. The resulting solution was refluxed for 3 hr., allowed to stand overnight, and refluxed for an additional hour. Twenty milliliters of water was added to the pre- cooled solution and the resulting suspension extracted with ether. The ether phase was washed with water, dried over sodium sulfate, and evaporated to dryness under reduced pressure. The resulting brown oil was extracted with petro- leum ether (t1.p. 30-60") and the estract evaporated to di,y ness. A light yellow oil resulted (14.2 mg.); A:: 5 . i 5 (cyclopentyl ketone), 6.05 (C=C), 8.05 p (acetate) and no hydroxyl present. The complex band a t 8.05 p similar to that of androsterone acetate indicated the presence of an axial acetate (3a,ja).8 The crude oil was dissolved in 4.0 ml. methanol containing 55 mg. of potassium carbonate dis- solved in 1.0 ml. of water and the mixture reflused for 2 hr. Water wan added to form a suspension which was ex- tracted with about 100 ml. of ether, the ether phase was washed with w-at,er, dried over sodium sulfat,e, and evapo- rated to dryness under reduced pressure. The colorless oil obtained (8.5 mg.) was chromatographed on a silica gel column and eluted with benzene and benzene-ethyl acetate mixtures. The 2.7-mg. sample of white amorphous ma- terial which \vas eluted with benzene gave 110 significant ultraviolet absorption in the region of 220-360 nip; A,",!: 5.75 (cyclopentyl ketone), Gp (isolated double bond), and no hydroxyl or acetate absorptions. Based on the infrared spectra and on reaction? carried out under similar conditions with epiandrosterones and allopregnane-3a-ol,11,20-dione7 which yielded the corresponding A*-elimination products, the compound is tentatively assigned the structure of A2-19- norandrostan-17-one (m.p. 115-121'). Elution with hen- gene-ethyl acetate 9: 1 and 6: 1 resulted in 3.7 mg. of white amorphous material which upon crystallization from are- tone-hexane yielded (11) colorless needles with the double melt 148O, 164.5-167"; [a]:,' $110, (c , 0.765 in chloro- form); 2.75 (OH), 5.75 (cyclopentyl C=O), 9.0, 9.35, 9.49, 9.65, 9.81, 10 p (axial OH).*

1 ~ - . r o r - 5 a - a n d r o s t a n e - ~ ~ , 1 7 p - ~ i o / (111) f r o m (11). A 2.1- mg. sample of I1 was t h o l v e d in 1.0 ml. of methanol con- taining 15 nig. of sodium borohydride. The solution was stirred overnight, n-ater as added, and the resulting sus- pension extracted with ether. The ether extract was washed with nat,er, dried over sodium sulfate, and evaporated to dryness. Chromatography on silica gel yielded 1.3 mg. of white amorphous material (111). Crystallization from ace-

( 5 ) All melting points are uncorrected. (6) D. Kupfer, E. Forchielli, and R. I. Dorfman, J . Ant.

(7) Yon ifr. Nagata, C. Tamm, and T. Reichstein, Helv.

(8) D. H. Barton, J . Chern. Soc., 1027 (1953). (9) ,J. Iriarte, G. Rosenkrans, and F. Sondheimer, J .

Cheni. Soc., 82, 1257 (1960).

Chinz. Actn. 42, 1399 (1959).

Ory. Cherri. 20, 342 (1055).

tone-hexane produced colorless needles, m.p. 191-193': [a]:" + 23.7 (c , 0.34 in chloroform); 2.90 (bonded OH), and 9.15, 9.40, 9.55, 9.90, 10.00 p (axial OH).8 Oxida- tion of the diol with c-hrornic acid in acetic acid produced a dione (Xi:; 5 80, 5.Si p ) identical to ail osidation product of lt)-noi-5a-androstane-3p, 170 diol and to an authentic sample of 5a-l~-iiorandrostane-B,17-dioiie.l0~'~

Acknowledgment. The authors wish to thank 3Ir. Donald W. Parsons for his excellent technical assistance.

THE \$-ORCESTER FOUNDATION FUR EXPERIMENTAL BIOLOGY

SHREWSBURY, MASS.

(10) C. Chen, Tetrahedron 3 ,43 (1958). (11) This compound was kindly supplitd by L)r. 1lih:i

Hayano.

11 -Oxygenated 17a-Acetoxy-9a-fluor0-6a- methyl- 1,4-pregnadiene-3,2O-diones

BARNEY J. 3IAGERLEIN AND FRED &G.4X

Received February 10, 1960

Acylation of Sa-fluoro-1 lp,l'icu-dihydroxy-(joc- methyl-1,4-pregnadiene-3,20-dione (I) lv2 with acc- tic anhydride-p-toluenesulfonic acid3 afforded 17a- acetoxy-9a- fluoro - 116 - hydroxy - 6a - methyl - 1,4- pregnadiene-3,20-dione (11) in 45% yield. The 11- keto analog I11 was obtained by the chromic acid oxidation of 11.

Endocrine assays of these compounds are sum- marized in Table I.

T.IBLE I CORTICOID A X D PROGESTATIOS 41. . A . S S , ~ T ~ ot'

COMPOUNDS I, 11, A X D 111

Anti- Inflammatory

Activity Glycogeri I'roges- (S Hydro- Deposition tational

Com- cortisone) (S Hydro- ( X Progrs- pound Rats cortisone) t erone)

I 1314 264 60 I1 1706 7 6 60-805

111 40

Conipound I1 is the only steroid tlescrilwd :L.S

effectively inhibiting both the C-3-E3 maninnary

(1) J. A. Hogg, 6th National Medicinal Chemistry Sym- posium, ACS, Madison, Wis., June 23, 1958.

(2) The registered trademark of the Upjohn Company, Kalamasoo, Mich., for 9a-fluoro-llj3,17a-dihydroxy-6a- methyl-1,4-pregnadiene-3,20-dione is Ox? lone.

(3) R. B. Turner, J . Am. Chem. Soc., 75, 3489 (1953). (4) R. 0. Stafford, A. Robert, S. C. Lyster, F. L. Schmidt,

and R. E. Ihlin, Proc. Sac. Exptl. Riol. Afed., 101, ti53 (1!15O).

1676 NOTES VOL. 25

adenocarcinoma in mice and the testosterone propionate-resistant mammary fibroadenoma in rats5 (see Table 11).

TABLE I1 TUMOR INHIBITION BY COMPOUND 11

yo Tumor Inhibitiona TP-

Dose, Resistantb C-3-H Compound mg./kg. (Rats) (Mice)

Prednisolone 15 0-20 100 Testosterone 5 0-15 0

Compound 15 72 96 propionate

I1

a Results derived from multiple assays, using 8-20 animals/group. The steroids were administered subcutane- ously in a CMC vehicle. E. M. Glenn, S. L. Richardson, and B. J. Bowman, Endocrinology, 64, 379 (1959).

EXPERIMENTAL^ 1 ?'cu-dcetox y-9a-fluoro-1 ip-hydroxy-6a-niethyl-1 ,.C-pregna-

diene-5,ZO-dione (11). A mixture of 5.0 g. of Sa-fluoro- 1 1p, 17~dihydroxy-6cu-methyl-1,4 - pregnadiene - 3,20 - diont: (I) in 625 ml. of glacial acetic acid, 125 ml. of acetic anhy- dride, and 2.0 g. of p-toluenesulfonic acid, monohydrate, was stirred vigorously a t 26" until solution was completed (about 8 hr.). The reaction mixture was poured into a large volume of water containing 1 kg. of potassium bicarbonate. The product recovered by filtration, after drying, melted at 205- 212' and constituted a quantitative yield. The crude prod- uct was recrystallized from ethyl acetate-Skellysolve B7 to afford 2.5 g. (45.0% yield) of 11, m.p. 225-228", [ a ] ~ + 49' (pyridine). The analytical sample, m.p. 230-232", [ a ] ~ + 50" (pyridine), mas prepared by recrystallization from the same solvents.

Anal. Calcd. for CzrHalFOs: C, 68.89; H, 7.47; F, 4.54. Found: C, 68.90; H, 7.56; F, 4.7.

17~-Acetoxy-9~~-Ruoro-6~~-methyl-l ,.C-pregnadiene-3, 11,60- trione (111). To a solution of 1 g. of I1 in 50 ml. of acetone, 0.5 ml. of chromic acid solutions was added with stirring. After 5 min. the excess oxidizing agent was destroyed by the addition of a few drops of methanol. The reaction mixture was concentrated under vacuum and the product isolated by partition between methylene dichloride-water. The residue obtained from the methylene dichloride fraction when recrystallized from ethyl acetate-Skellysolve B weighed 300 mg. (30%) and melted at. 273-275". Recrystal- lization from the same solvents gave an analytical sample, m.p. 277-2785".

,477422. Calcd. for C ~ ~ H W F O ~ : C, 69.21; H, 7.07; F, 4.56. Found: C, 69.44; H, 7.36; F, 4.5.

THE UPJOHN Co. KALAMAZOO, MWH.

(5) Presented bv E. &I. Glenn, S. L. Richardson, B. J. Bowman, and S. C. Lyster a t CCNSC Symposium titled, "Biologic Activities of Steroids in Relation to Cancer," Vergennes, Vt., Sept. 27-Oct. 2, 1959. (Abstracts of papers to be published.)

(6) The authors are indebted to G. E. VandenBerg of these laboratories for assistance in the preparation of these compounds.

(7) A saturated hydrocarbon fraction, b.p. 6@71 O .

(8) A. Bowers, T. G. Halsall, E. R. H. Jones, and A. J. Lemin, J . Chem. SOC., 2548 (1953).

The Preparation of 16-Methyl-A16-steroids Containing Ring C Substituents

SEYMOUR BERNSTEIN AND J. P. JOSEPH

Received March 9, 1960

Research in this laboratory on C14-substituted steroids1 has been extended to include methyl substituents. In view of the recent publications,2 especially that by Slates and Wendlerj2g on C16-

methyl steroids we wish to report here on our work in this area. This note describes the preparation of 16-methyl-A"hteroids which contain substituents in the C-ring, in particular, on 21-acetoxy-9a- fluoro - 1 lp-hydroxy- 16-methy1-4,16-pregnadiene-3 ,- 20-dione (VII) .

Following the procedure of Wettstein3 21-ace- toxy-4,9( ll), 16-pregnatriene-3,20-dione4 (Ia) on reaction with excess diazomethane gave 21- acetoxy-16a, 17a- [3,1-( l-pyrazolino) ] - 4,9 (1 1) - preg- nadiene-3,20-dione (IIa).5 A band attributable to --N=N-stretchingb was observed a t 1565 cm. -l

in this and all the other pyrazolino-steroids herein prepared. Thermal decomposition of I Ia readily afforded 21-acetoxy-16-methyl-4,9( 11),16-pregna- triene-3,20-dione (IIIa).

N-bromoacetamide-perchloric acid treatment of 21-acetoxy-16a,l7a-[3,1- (1 - pyrazolino)] - 4,9(11) - bromohydrin IV. This was converted, without further purification, in refluxing methanolic potas- sium acetate directly into 2l-acetoxy-l6a,l7a- [3,1-( l-pyrazolin0)]-9/3,1 lp-epoxy-4-pregnane-3,20- dione(V).

( 1 ) S. Bernstein, R. H. Lenhard, W. S. Allen, M. Heller, R. Littell, S. M. Stolar, L. I. Feldman, R. H. Blank, J . Am. Chem. Soc., 78, 5693 (1966), and subsequent papers.

(2) (a) G. E. Arth, D. B. Johnston, J. Fried, W. W. Spooncer, D. R. Hoff, and L. H. Sarett, J . Am. Chem. SOC., 80, 3160 (1958). (b) G. E. Arth, J. Fried, D. B. Johnston, D. R. Hoff, L. H. Sarett, R. H. Silber, H. C. Stoerk, and C. A. Winter, J . Am. Chem. SOC., 80, 3161 (1958). (c) D. Taub, R. D. Hoffsommer, H. L. Slates, and N. L. Wendler, J . Am. Chem. SOC., 80, 4435 (1958). (d) E. P. Oliveto, R. Rausser, A. L. Nussbaum, W. Gebert, E. B. Hershberg, S. Tolksdorf, M. Eisler, P. L. Perlman, and M. 11. Pechet, J . Am. Chem. Soc., 80, 4428 (1958). (e) E. P. Oliveto, R. Rausser, L. Weber, A. L. Xussbaum, W. Gebert, C. T. Coniglio, E. B. Hershberg, S. Tolksdorf, A I . Eisler, P. L. Perlman, and M. M. Pechet, J . Am. Chena. SOC., 80, 4431 (1958). (f) E. P. Oliveto, R. Rausser, H. L. Herzog, E. B. Hershberg, S. Tolksdorf, M. Eisler, P. L. I'erlman, and M. A I . Pechet, J. Am. C h m . SOC., 80, 6687 (1958). (g) H. L. Slates and N. L. Wendler, J . .4m. Chem. Soc., 81, 5472 (1959).

(3) A. Wettstein, Helv. Chim. Acta, 27, 1803 (1944). (4) W. S. Allen and S. Bernstein, J . A m . Chem. SOC., 78,

1909 (1956). (5) This system of nomenclature for pyrazoline deriva-

tives is according to that employed by G. P. Mueller and B. Riegel, J . ,4m. Chem. SOC., 76, 3686 (1954) for similar compounds.

(6) R. N. Jones and C. Ehndorfy, Techniques of Organic Chemistry, 9, 545 (1956).